Document Detail


Measurement of thyroid stimulating immunoglobulins using a novel thyroid stimulating hormone receptor-guanine nucleotide-binding protein, (GNAS) fusion bioassay.
MedLine Citation:
PMID:  23039881     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Hyperthyroidism, defined by overproduction of thyroid hormones, has a 2-3% prevalence in the population. The most common form of hyperthyroidism is Graves' disease. A diagnostic biomarker for Graves' disease is the presence of immunoglobulins which bind to, and stimulate, the thyroid stimulating hormone receptor (TSHR), a G-protein coupled receptor (GPCR). We hypothesized that the ectopically expressed TSHR gene in a thyroid stimulating immunoglobulin (TSI) assay could be engineered to increase the accumulation of the GPCR pathway second messenger, cyclic AMP (cAMP), the molecule measured in the assay as a marker for pathway activation. An ectopically expressing TSHR-mutant guanine nucleotide-binding protein, (GNAS) Chinese hamster ovary (CHO) cell clone was constructed using standard molecular biology techniques. After incubation of the new clone with sera containing various levels of TSI, GPCR pathway activation was then quantified by measuring cAMP accumulation in the clone. The clone, together with a NaCl-free cell assay buffer containing 5% polyethylene glycol (PEG)6000, was tested against 56 Graves' patients, 27 toxic thyroid nodule patients and 119 normal patients. Using receiver operating characteristic analysis, when comparing normal with Graves' sera, the assay yielded a sensitivity of 93%, a specificity of 99% and an efficiency of 98%. Total complex precision (within-run, across runs and across days), presented as a percentage coefficient of variation, was found to be 7·8, 8·7 and 7·6% for low, medium and high TSI responding serum, respectively. We conclude that the performance of the new TSI assay provides sensitive detection of TSI, allowing for accurate, early detection of Graves' disease.
Authors:
M Pierce; R Sandrock; G Gillespie; A W Meikle
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Clinical and experimental immunology     Volume:  170     ISSN:  1365-2249     ISO Abbreviation:  Clin. Exp. Immunol.     Publication Date:  2012 Nov 
Date Detail:
Created Date:  2012-10-08     Completed Date:  2013-03-18     Revised Date:  2013-11-05    
Medline Journal Info:
Nlm Unique ID:  0057202     Medline TA:  Clin Exp Immunol     Country:  England    
Other Details:
Languages:  eng     Pagination:  115-21     Citation Subset:  IM    
Copyright Information:
© 2012 ARUP Institute for Clinical and Experimental Pathology. Clinical and Experimental Immunology © 2012 British Society for Immunology.
Affiliation:
ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT 84108-1221, USA. michael.l.pierce@aruplab.com
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MeSH Terms
Descriptor/Qualifier:
Animals
Biological Assay / methods*
CHO Cells
Cells, Cultured
Cricetinae
Cyclic AMP / genetics,  metabolism
GTP-Binding Proteins / chemistry*,  genetics,  metabolism
Graves Disease / blood,  genetics,  metabolism
Humans
Hyperthyroidism / blood,  genetics,  metabolism
Immunoglobulins, Thyroid-Stimulating / blood*,  chemistry*,  metabolism
Polyethylene Glycols / chemistry
Receptors, G-Protein-Coupled / chemistry*,  genetics,  metabolism
Receptors, Thyrotropin / chemistry*,  genetics,  metabolism
Sensitivity and Specificity
Thyroid Gland / metabolism
Chemical
Reg. No./Substance:
0/Immunoglobulins, Thyroid-Stimulating; 0/Polyethylene Glycols; 0/Receptors, G-Protein-Coupled; 0/Receptors, Thyrotropin; 60-92-4/Cyclic AMP; EC 3.6.1.-/GTP-Binding Proteins
Comments/Corrections

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